CFTR involvement in nasal potential differences in mice and pigs studied using a thiazolidinone CFTR inhibitor

Nasal potential difference (PD) measurements have been used to demonstrate defective CFTR function in cystic fibrosis (CF) and to evaluate potential CF therapies. We used the selective thiazolidinone CFTR inhibitor CFTR(inh)-172 to define the involvement of CFTR in nasal PD changes in mice and pigs. In normal mice infused intranasally with a physiological saline solution containing amiloride, nasal PD was -4.7 +/- 0.7 mV, hyperpolarizing by 15 +/- 1 mV after a low-Cl- solution, and a further 3.9 +/- 0.5 mV after forskolin. CFTR(inh)-172 produced 1.1 +/- 0.9- and 4.3 +/- 0.7-mV depolarizations when added after low Cl- and forskolin, respectively. Systemically administered CFTR(inh)-172 reduced the forskolin-induced hyperpolarization from 4.7 +/- 0.4 to 0.9 +/- 0.1 mV but did not reduce the low Cl(-)-induced hyperpolarization. Nasal PD was -12 +/- 1 mV in CF mice after amiloride, changing by <0.5 mV after low Cl- or forskolin. In pigs, nasal PD was -14 +/- 3 mV after amiloride, hyperpolarizing by 13 +/- 2 mV after low Cl- and a further 9 +/- 1 mV after forskolin. CFTR(inh)-172 and glibenclamide did not affect nasal PD in pigs. Our results suggest that cAMP-dependent nasal PDs in mice primarily involve CFTR-mediated Cl- conductance, whereas cAMP-independent PDs are produced by a different, but CFTR-dependent, Cl- channel. In pigs, CFTR may not be responsible for Cl- channel-dependent nasal PDs. These results have important implications for interpreting nasal PDs in terms of CFTR function in animal models of CFTR activation and inhibition.     

Introducing weights to life cycle sustainability assessment—how do decision-makers weight sustainability dimensions?

Purpose

Decisions based on life cycle sustainability assessment (LCSA) pose a multi-criteria decision issue, as impacts on the three different sustainability dimensions have to be considered which themselves are often measured through several indicators. To support decision-making at companies, a method to interpret multi-criteria assessment and emerging trade-offs would be beneficial. This research aims at enabling decision-making within LCSA by introducing weights to the sustainability dimensions.

Methods

To derive weights, 54 decision-makers of different functions at a German automotive company were asked via limit conjoint analysis how they ranked the economic, environmental, and social performance of a vehicle component. Results were evaluated for the entire sample and by functional clusters. Additionally, sustainability respondents, i.e., respondents that dealt with sustainability in their daily business, were contrasted with non-sustainability respondents. As a last step, the impact of outliers was determined. From this analysis, practical implications for ensuring company-optimal decision-making in regard to product sustainability were derived.

Results and discussion

The results showed a large spread in weighting without clear clustering. On average, all sustainability dimensions were considered almost equally important: the economic dimension tallied at 33.5%, the environmental at 35.2%, and the social at 31.2%. Results were robust as adjusting for outliers changed weights on average by less than 10%. Results by function showed low consistency within clusters hinting that weighting was more of a personal than a functional issue. Sustainability respondents weighted the social before the environmental and economic dimension while non-sustainability respondents put the economic before the other two dimensions. Provided that the results of this research could be generalized, the retrieved weighting set was seen as a good way to introduce weights into an operationalized LCSA framework as it represented the quantification of the already existing decision process. Therefore, the acceptance of this weighting set within the respective company was expected to be increased.

Conclusions

It could be shown that conjoint analysis enabled decision-making within LCSA by introducing weights to solve a multi-criteria decision issue. Furthermore, implications for practitioners could be derived to ensure company-optimal decision-making related to product sustainability. Future research should look at expanding the sample size and geographical scope as well as investigating the weighting of indicators within sustainability dimensions and the drivers that influence personal decision-making in regard to weighting sustainability dimensions.

     

The implementation of organizational LCA to internally manage the environmental impacts of a broad product portfolio: an example for a cosmetics,fragrances, and toiletry provider

Purpose

This paper presents the implementation of O-LCA by a Brazilian cosmetics manufacturer. The case study was developed within the framework of the road testing of the “Guidance on organizational LCA” of the UNEP/SETAC Life Cycle Initiative. The aim is to illustrate methodological choices and implementation challenges encountered by the company, i.e., related to the broad product portfolio. The study demonstrates that O-LCA allows quantifying and managing environmental impacts throughout global supply chains and for every individual product.

Methods

O-LCA provides the methodological framework for applying LCA to organizations, and a set of application options based on the structure and experience of organizations. The reporting organization is NATURA Brazil in 2013. The 2600 products in the portfolio are modeled in this first exercise of the company through the bestsellers at each of its ten product category groups. A hybrid approach is considered for data collection: top-down approach for modeling corporate activities and bottom-up approach for upstream and downstream life cycle phases. The data sources are NATURA’s recordings, data gathered from suppliers, estimates from mass and energy balances, and life cycle inventory databases. The approach to acquire direct data or use life cycle databases depends on the representativeness of each raw material or packaging.

Results and discussion

The results show that major impacts could be detected during use phase that demands water and energy to use rinse-off products (the use phase of NATURA’s products contributed over 41% to most impact categories), and in the supply chain, and generated during the obtaining of plant origin ingredients and materials for packaging. Overall, the whole NATURA had in 2013 a potential impact on climate change of 1.4 million tonnes of CO₂ eq, a natural land transformation of 1.3 million m², and a fossil depletion of 0.23 million tonnes of oil eq, among other impacts. Apart from the results at the organizational level, individual results for product bestsellers were calculated and are presented here.

Conclusions

The study confirmed the applicability of the O-LCA model at NATURA, addressed operational issues related to broad product portfolios, considering several dimensions such as data quality and availability, LCA software, and data management. Despite NATURA’s existing practices and previous knowledge in modeling environmental impacts of products and corporate activities, managing the large amount of data involved prove being a complex task. The company identified gaps and opportunities able to guide future method implementation and LCA-based management.

     

Cytokinesis in yeast meiosis depends on the regulated removal of Ssp1p from the prospore membrane

Intracellular budding is a developmentally regulated type of cell division common to many fungi and protists. In Saccaromyces cerevisiae, intracellular budding requires the de novo assembly of membranes, the prospore membranes (PSMs) and occurs during spore formation in meiosis. Ssp1p is a sporulation-specific protein that has previously been shown to localize to secretory vesicles and to recruit the leading edge protein coat (LEP coat) proteins to the opening of the PSM. Here, we show that Ssp1p is a multidomain protein with distinct domains important for PI(4,5)P(2) binding, binding to secretory vesicles and inhibition of vesicle fusion, interaction with LEP coat components and that it is subject to sumoylation and degradation. We found non-essential roles for Ssp1p on the level of vesicle transport and an essential function of Ssp1p to regulate the opening of the PSM. Together, our results indicate that Ssp1p has a domain architecture that resembles to some extent the septin class of proteins, and that the regulated removal of Ssp1p from the PSM is the major step underlying cytokinesis in yeast sporulation.     

To fear or not to fear: what was the question? A potential role for Ras-GRF in memory

Learning, making memories, and forgetting are thought to require changes in the strengths of connections between neurons. Such changes in synaptic strength occur in two phases: an early phase that is likely mediated by covalent modifications to existing proteins, and a delayed phase that depends on new gene expression and protein synthesis. However, the biochemical mechanisms by which neuronal activity leads to changes in synaptic strength are poorly understood. Recently, it has been shown that animals that lack Ras guanine nucleotide releasing factor (Ras-GRF), a Ca²⁺-dependent activator of the small GTP-binding protein, Ras, do not learn fear responses normally, although other types of learning appear normal. These animals show defects in the delayed phase of memory formation within the neuronal circuit that mediates fear conditioning. This paper suggests that Ras-GRF couples synaptic activity to the molecular mechanisms that consolidate changes in synaptic strength within specific neuronal circuits.^(1–3) BioEssays 20 :691–695, 1998. © 1998 John Wiley & Son, Inc.     

Analysis of the potential for a comprehensive approach towards LCA and EMS in Japan

Sustainable development can only be achieved if industry adoptsboth product related and organisation related environmental management tools, such as Life Cycle Assessment (LCA) and Environmental Management Systems (EMS). In Japan, EMS (ISO 14001) is more widely applied than LCA (ISO 14040). Therefore,one means by which Japanese industries could be motivated to adopt and use LCA is to relate LCA-activities to the policies and instruments of ISO 14001. The potential of such a comprehensive approach was analysed by a survey of 270 Japanese enterprises (response rate 45%). The results indicate that 19% of the responding representatives had responsibilities for both LCA and EMS, while the remaining only work in one of both fields. A statement in the company’s/ plant’s Environmental Policy of ISO 14001, stating that LCA is to be used as part of the EMS, was found in 42% of all companies. A surprising number (39%) either already use, or plan to use, LCA and EMS as combinated/integrated tools. A strong argument for the establishment of a comprehensive approach can be seen in the perception of the usefulness of LCA, which was rated significantly higher in companies that acknowledged the complementary potential of LCA and EMS.     

Proteolysis of Mutant Huntingtin Produces an Exon 1 Fragment That Accumulates as an Aggregated Protein in Neuronal Nuclei in Huntington Disease

Huntingtin proteolysis has been implicated in the molecular pathogenesis of Huntington disease (HD). Despite an intense effort, the identity of the pathogenic smallest N-terminal fragment has not been determined. Using a panel of anti-huntingtin antibodies, we employed an unbiased approach to generate proteolytic cleavage maps of mutant and wild-type huntingtin in the HdhQ150 knock-in mouse model of HD. We identified 14 prominent N-terminal fragments, which, in addition to the full-length protein, can be readily detected in cytoplasmic but not nuclear fractions. These fragments were detected at all ages and are not a consequence of the pathogenic process. We demonstrated that the smallest fragment is an exon 1 huntingtin protein, known to contain a potent nuclear export signal. Prior to the onset of behavioral phenotypes, the exon 1 protein, and possibly other small fragments, accumulate in neuronal nuclei in the form of a detergent insoluble complex, visualized as diffuse granular nuclear staining in tissue sections. This methodology can be used to validate the inhibition of specific proteases as therapeutic targets for HD by pharmacological or genetic approaches.     

Environmental performance of building materials: life cycle assessment of a typical Sicilian marble

Background, aim, and scope  

The building sector is strategically important for achieving sustainability. Therefore, the improvement of energy and environmental performances are relevant targets because precious building materials such as marble have a significant impact on the environment. The aim of this paper is an analysis of a typical Sicilian marble (Perlato di Sicilia) to evaluate its energy and environmental performance. Marble plays an important role in the economy of Italy and has a global market share of 58% in terms of exports. For the main production areas of marble, relevant environmental performance data are missing except for one region (Tuscany—Massa e Carrara province). Perlato di Sicilia, the main marble of Custonaci (Sicily), has never been analyzed previously.